Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection

Research output: Contribution to conference without publisher/journalPosterResearchpeer-review

132 Downloads (Pure)

Abstract

Human Milk Oligosaccharides (HMOs) are composed of 5 different monosaccharides: D-glucose, D-galactose, L-fucose, N-acetylneuraminic acid and N-acetylglucosamine. Approximately 200 unique structures have been identified, ranging in the degree of polymerization from 3 to 22. The diversity among different mothers is considerable, ranging from as few as 23 and up to 130 different oligosaccharides. HMOs are known to be beneficial for infant health and development, and have received a lot of attention in recent years (Bode & Jantscher-Krenn 2012).
Syntheses of this unique family of lactose-based molecules necessitates analysis methods that can provide separation and quantification of the common structural constituents mentioned, as well as the disaccharide lactose (Galβ1-4Glc) and oligosaccharides such as lacto-N-triose II (GlcNAcβ1-3Galβ1-4Glc), lacto-N-tetraose (Galβ1-3GlcNAcβ1-3Galβ1-4Glc) and lacto-N-neotetraose (Galβ1-4GlcNAcβ1-3Galβ1-4Glc), among others.
High-performance anion-exchange chromatography (HPAE) with pulsed amperometric detection (PAD) is an analysis method highly suited for carbohydrates. HPAE with alkaline eluents results in retention of neutral carbohydrates depending on the number of charged group in the molecule, pH and concentration of competing anions, while the PAD has sensitivity for carbohydrates in the pmol-range (Lee 1990).
As a basis for the development and optimisation of HPAE elution methods, the parameter space was investigated in terms of eluent concentrations and temperature, using a CarboPac PA1 column. Specifically, the retention times and separations of various mono- and oligosaccharides, as depending on the eluent concentrations of NaOH and NaOAc, was studied. The investigated monosaccharides and several of the disaccharides exhibited maximum retention at distinct concentrations of NaOH. In addition, the different species showed varying relations between eluent composition and retention time, resulting in varying separation and even changes in elution order with different elution conditions.

References:
Bode L, Jantscher-Krenn E. (2012) Adv. Nutr. 3: 383S
Lee YC. (1990) Anal. Biotechnol. 189: 151
Original languageEnglish
Publication date26 May 2016
Number of pages1
Publication statusPublished - 26 May 2016
Event11th Danish Conference on Biotechnology and Molecular Biology: Glycobiology and Carbohydrate Biotechnology - Hotel Munkebjerg, Vejle, Denmark
Duration: 26 May 201627 May 2016
http://danishbiotechsociety.org/previous-conferences-events/2016-dcb11/

Conference

Conference11th Danish Conference on Biotechnology and Molecular Biology
LocationHotel Munkebjerg
CountryDenmark
CityVejle
Period26/05/201627/05/2016
Internet address

Fingerprint

Chromatography
Oligosaccharides
Anions
Monosaccharides
Disaccharides
Carbohydrates
Lactose
Molecules
Fucose
Acetylglucosamine
N-Acetylneuraminic Acid
Galactose
Polymerization
Human Milk
Health
Glucose
Chemical analysis
Temperature
lacto-N-neotetraose

Keywords

  • Human milk oligosaccharides
  • Analysis methods
  • Anion-exchange chromatography
  • Pulsed amperometric detection
  • Method development

Cite this

Lie, A., & Pedersen, L. H. (2016). Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection. Poster session presented at 11th Danish Conference on Biotechnology and Molecular Biology, Vejle, Denmark.
Lie, Aleksander ; Pedersen, Lars Haastrup. / Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection. Poster session presented at 11th Danish Conference on Biotechnology and Molecular Biology, Vejle, Denmark.1 p.
@conference{a9ce79062b4346f9bc4a0cfc72928d57,
title = "Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection",
abstract = "Human Milk Oligosaccharides (HMOs) are composed of 5 different monosaccharides: D-glucose, D-galactose, L-fucose, N-acetylneuraminic acid and N-acetylglucosamine. Approximately 200 unique structures have been identified, ranging in the degree of polymerization from 3 to 22. The diversity among different mothers is considerable, ranging from as few as 23 and up to 130 different oligosaccharides. HMOs are known to be beneficial for infant health and development, and have received a lot of attention in recent years (Bode & Jantscher-Krenn 2012).Syntheses of this unique family of lactose-based molecules necessitates analysis methods that can provide separation and quantification of the common structural constituents mentioned, as well as the disaccharide lactose (Galβ1-4Glc) and oligosaccharides such as lacto-N-triose II (GlcNAcβ1-3Galβ1-4Glc), lacto-N-tetraose (Galβ1-3GlcNAcβ1-3Galβ1-4Glc) and lacto-N-neotetraose (Galβ1-4GlcNAcβ1-3Galβ1-4Glc), among others.High-performance anion-exchange chromatography (HPAE) with pulsed amperometric detection (PAD) is an analysis method highly suited for carbohydrates. HPAE with alkaline eluents results in retention of neutral carbohydrates depending on the number of charged group in the molecule, pH and concentration of competing anions, while the PAD has sensitivity for carbohydrates in the pmol-range (Lee 1990).As a basis for the development and optimisation of HPAE elution methods, the parameter space was investigated in terms of eluent concentrations and temperature, using a CarboPac PA1 column. Specifically, the retention times and separations of various mono- and oligosaccharides, as depending on the eluent concentrations of NaOH and NaOAc, was studied. The investigated monosaccharides and several of the disaccharides exhibited maximum retention at distinct concentrations of NaOH. In addition, the different species showed varying relations between eluent composition and retention time, resulting in varying separation and even changes in elution order with different elution conditions.References:Bode L, Jantscher-Krenn E. (2012) Adv. Nutr. 3: 383SLee YC. (1990) Anal. Biotechnol. 189: 151",
keywords = "Human milk oligosaccharides, Analysis methods, Anion-exchange chromatography, Pulsed amperometric detection, Method development",
author = "Aleksander Lie and Pedersen, {Lars Haastrup}",
year = "2016",
month = "5",
day = "26",
language = "English",
note = "11th Danish Conference on Biotechnology and Molecular Biology : Glycobiology and Carbohydrate Biotechnology, DCB11 ; Conference date: 26-05-2016 Through 27-05-2016",
url = "http://danishbiotechsociety.org/previous-conferences-events/2016-dcb11/",

}

Lie, A & Pedersen, LH 2016, 'Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection' 11th Danish Conference on Biotechnology and Molecular Biology, Vejle, Denmark, 26/05/2016 - 27/05/2016, .

Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection. / Lie, Aleksander; Pedersen, Lars Haastrup.

2016. Poster session presented at 11th Danish Conference on Biotechnology and Molecular Biology, Vejle, Denmark.

Research output: Contribution to conference without publisher/journalPosterResearchpeer-review

TY - CONF

T1 - Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection

AU - Lie, Aleksander

AU - Pedersen, Lars Haastrup

PY - 2016/5/26

Y1 - 2016/5/26

N2 - Human Milk Oligosaccharides (HMOs) are composed of 5 different monosaccharides: D-glucose, D-galactose, L-fucose, N-acetylneuraminic acid and N-acetylglucosamine. Approximately 200 unique structures have been identified, ranging in the degree of polymerization from 3 to 22. The diversity among different mothers is considerable, ranging from as few as 23 and up to 130 different oligosaccharides. HMOs are known to be beneficial for infant health and development, and have received a lot of attention in recent years (Bode & Jantscher-Krenn 2012).Syntheses of this unique family of lactose-based molecules necessitates analysis methods that can provide separation and quantification of the common structural constituents mentioned, as well as the disaccharide lactose (Galβ1-4Glc) and oligosaccharides such as lacto-N-triose II (GlcNAcβ1-3Galβ1-4Glc), lacto-N-tetraose (Galβ1-3GlcNAcβ1-3Galβ1-4Glc) and lacto-N-neotetraose (Galβ1-4GlcNAcβ1-3Galβ1-4Glc), among others.High-performance anion-exchange chromatography (HPAE) with pulsed amperometric detection (PAD) is an analysis method highly suited for carbohydrates. HPAE with alkaline eluents results in retention of neutral carbohydrates depending on the number of charged group in the molecule, pH and concentration of competing anions, while the PAD has sensitivity for carbohydrates in the pmol-range (Lee 1990).As a basis for the development and optimisation of HPAE elution methods, the parameter space was investigated in terms of eluent concentrations and temperature, using a CarboPac PA1 column. Specifically, the retention times and separations of various mono- and oligosaccharides, as depending on the eluent concentrations of NaOH and NaOAc, was studied. The investigated monosaccharides and several of the disaccharides exhibited maximum retention at distinct concentrations of NaOH. In addition, the different species showed varying relations between eluent composition and retention time, resulting in varying separation and even changes in elution order with different elution conditions.References:Bode L, Jantscher-Krenn E. (2012) Adv. Nutr. 3: 383SLee YC. (1990) Anal. Biotechnol. 189: 151

AB - Human Milk Oligosaccharides (HMOs) are composed of 5 different monosaccharides: D-glucose, D-galactose, L-fucose, N-acetylneuraminic acid and N-acetylglucosamine. Approximately 200 unique structures have been identified, ranging in the degree of polymerization from 3 to 22. The diversity among different mothers is considerable, ranging from as few as 23 and up to 130 different oligosaccharides. HMOs are known to be beneficial for infant health and development, and have received a lot of attention in recent years (Bode & Jantscher-Krenn 2012).Syntheses of this unique family of lactose-based molecules necessitates analysis methods that can provide separation and quantification of the common structural constituents mentioned, as well as the disaccharide lactose (Galβ1-4Glc) and oligosaccharides such as lacto-N-triose II (GlcNAcβ1-3Galβ1-4Glc), lacto-N-tetraose (Galβ1-3GlcNAcβ1-3Galβ1-4Glc) and lacto-N-neotetraose (Galβ1-4GlcNAcβ1-3Galβ1-4Glc), among others.High-performance anion-exchange chromatography (HPAE) with pulsed amperometric detection (PAD) is an analysis method highly suited for carbohydrates. HPAE with alkaline eluents results in retention of neutral carbohydrates depending on the number of charged group in the molecule, pH and concentration of competing anions, while the PAD has sensitivity for carbohydrates in the pmol-range (Lee 1990).As a basis for the development and optimisation of HPAE elution methods, the parameter space was investigated in terms of eluent concentrations and temperature, using a CarboPac PA1 column. Specifically, the retention times and separations of various mono- and oligosaccharides, as depending on the eluent concentrations of NaOH and NaOAc, was studied. The investigated monosaccharides and several of the disaccharides exhibited maximum retention at distinct concentrations of NaOH. In addition, the different species showed varying relations between eluent composition and retention time, resulting in varying separation and even changes in elution order with different elution conditions.References:Bode L, Jantscher-Krenn E. (2012) Adv. Nutr. 3: 383SLee YC. (1990) Anal. Biotechnol. 189: 151

KW - Human milk oligosaccharides

KW - Analysis methods

KW - Anion-exchange chromatography

KW - Pulsed amperometric detection

KW - Method development

M3 - Poster

ER -

Lie A, Pedersen LH. Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection. 2016. Poster session presented at 11th Danish Conference on Biotechnology and Molecular Biology, Vejle, Denmark.